Summary and Results
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| Figure 1 |
Our gel successfully separated DNA fragments creating unique banding patterns for each restriction enzyme used. Fig 1 illustrates our final stained gel showing separation of DNA fragments and digital analysis of distance traveled on left. We used the HindiIII bands (bottom well) as a molecular
weight ruler or marker as we knew the size of these fragments. We then estimated the length of unknown DNA band sizes by constructing a a standard curve( figure 2) based upon the measurements obtained from the known DNA HindIII bands. Using Excel's trend line function, we found the relationship between fragment size and distance traveled to be
fragment size in base pairs = 54892e^(-0.172*distance in mm)
This regression line has a high coefficient of determination of 0.9975 (between 0 and 1.0, used to describe how well a regression line fits a set of data).
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| Figure 2 |
Procedure
Gels are placed in electrophoresis chamber which is filled with buffer which helps current flow.
DNA digests are loaded into wells at negative end of the gel.
From left: uncut lamda DNA, Pstl restriction digest, EcoRI restriction digest
HindiIII restriction digest
running the gels
the loading dye moved across the gel and can be observed easily
pouring the staining solution over the gel to help visualize the DNA bands.
destaining the gels before analysis
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